...cyanobacteria, synthetic inverters in E. coli, and biosensors in E. coli: Čelešnik, Helena, et al. "Biosafety...tune your polycistronic E. coli expression constructs. Some applications of plasmids from this kit include...epitope tags. Different plasmids in the kit can be used to purify proteins from E. coli, Bacillus subtilis,...curated collections of plasmids as kits with greatly reduced costs per plasmid. We generally create kits...just the tool you need. The plasmids in this portion of the BIOFAB Plasmid Set contain terminators of ...populations. This kit consists of a 48 plasmid adapter library, with each plasmid carrying a unique barcoded Illumina-M13F...synthetic biology or protein purification studies in E. coli. 800 and 1900 Series If you’re expressing two genes...

...amylovora, E. carotovora subsp. Carotovora, E. herbicola, E. rubrifaciens, E. stewartii, Escherichia coli, Klebsiella...Addgene blog Plasmids 101: Origin of Replication Plasmids 101: What is a Plasmid?  Plasmids 101: Transformation...host range in plasmids? Not all plasmids were created equally. Only a handful of plasmids have been identified...Conveniently, these broad host range plasmids all replicate in E. coli, which is the most commonly used bacteria...broad host-range IncQ plasmids. Plasmid, 62(2), 57–70. https://doi.org/10.1016/j.plasmid.2009.05.001  Meyer...define plasmid host range, identify a few of the features that confer broad host range in plasmids in nature...recombinant vectors.  A plasmid’s host range is determined by the different hosts that a plasmid can transfer to...

... the plasmids from Addgene is have the students transform them into different strains of E. coli,” says... Find fluorescent protein plasmids at Addgene Find chromoprotein plasmids at Addgene Check out additional...decided to add a bit of fun. He mixed together 10 E. coli strains producing different pigments, and tasked...ambient light. Expressing these chromoproteins in E. coli would make it easy to distinguish colonies from...other species.  When he found the chromoprotein plasmids available from Addgene, he knew it would be a ...plate skills? Find the chromoprotein expressing plasmids here, and check out our streak plate protocol ...

... some E. coli strains have natural antibiotic resistances, so make sure your plasmid and E. coli strain... much easier it makes our lives. Plasmid transformation into E. coli is a fairly inefficient process– ... presence of a plasmid is disadvantageous from the bacterium's perspective – a plasmid-containing cell...to maintain the plasmid. Adding an antibiotic resistance gene to the plasmid solves both problems at once...Biology Reference Pages View our Plasmid Protocols Find Your Perfect Plasmid Backbone ... are compatible! Check out this list of common E. coli genotypes and their natural resistances.  Additional... which cells successfully received the correct plasmid, scientists would spend hours to days trying find...

...dsRNA-expressing E. coli that are mixed with liver paste. Prior to mixing with the liver paste, these E. coli are ...experiments in planarians with their recent plasmid deposit. These plasmids enable two main types of experiments... cloned into plasmids to investigate for each of these planarian species. These plasmids all have a PR-T4...insert the cDNA into the desired plasmid backbone. The Sánchez lab plasmids allow students and new labs to...Resources at Addgene.org Find plasmids from the Sánchez lab Find plasmids for other developmental models...of T7 polymerase encoded in the E. coli genome (usually controlled by IPTG). For this IPTG-induced expression...expression to work, students have to use a commercial E. coli strain that is designated as DE3. For Phagocata...

...expressing E. coli aided in ultrasound imaging of the mouse gastrointestinal tract. The E. coli strain Nissle...Addgene.org Find plasmids for in vivo imaging with fluorescent proteins Browse luciferase plasmids Find fluorescent...created two constructs optimized for expression in E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2...20 hours of growth following acoustic collapse E. coli re-express arg1 gas vesicles and restore their ...acoustic reporters for your research? arg1 and arg2 plasmids are available from Addgene! References 1 Bourdeau...

...   Fig. 1: Expressing bARGSer in E. coli. (a) Plasmid diagram of pBAD-bARGSer-AxeTxe. (b) Methods...that the plasmid could be maintained in the absence of antibiotics, resulting in the plasmid pBAD-bARGSer-AxeTxe... 1248 ng of CMV-gvpA-IRES-BFP plasmid and 954 ng CMV-gvpNJKFGWV plasmid into 70%-confluent well of HEK...highest levels of nonlinear acoustic contrast in E. coli. We deleted an unannotated hypothetical protein...bARGSer with several inducible expression systems in E. coli. The arabinose-inducible promoter pBAD and a low-copy...pBAD-bARGSer-AxeTxe, gas vesicle expression in E. coli is easy: simply grow cells in Lennox LB (10 g/L...expression in vivo is also straightforward: after the E. coli have colonized tumors, inject L-arabinose intraperitoneally...

...bacteria Before looking at an example plasmid, let's review how E. coli normally replicate their chromosomal...maintaining a stable concentration of plasmid. Similar to replication in E. coli, regulation primarily occurs ... to stringently regulate plasmid replication. For example, the plasmid ColIb-P9 encodes the protein RepZ...replication. In general, these plasmids tend to be low copy number. Conversely, plasmids that can initiate DNA ...growth against sustainability. Plasmid DNA replication and regulation Plasmids must also strike a balance:...a given plasmid has been reached, negative feedback circuits reduce replication of the plasmid to balance...balance the total plasmid number.  Let’s consider pSC101. This plasmid is stringently controlled and has...

...Split-BioID plasmids Find CRISPR plasmids for labeling genomic regions Find CRISPR plasmids for purifying...interaction detection (RaPID) method. RaPID borrows the E. coli biotin ligase BirA* from BioID and allows a researcher...interactions. Both methods rely on the promiscuous E. coli biotin ligase, BirA*, to biotinylate proteins that...subtilis. BASU was  compared head-to-head with the E. coli and BioID2 BirA*s in the context of RaPID for their... no detectable signal at the same time point. E. coli’s BirA* was tested at its optimum 18 hour time point...still generated a >30-fold enrichment compared to E. coli’s BirA*. Faster labeling kinetics also means that...question that RaPID could help answer? The RaPID plasmids are available from Addgene! References Ramanathan...

...if you're transforming a plasmid with ampicillin resistance.  After a plasmid is introduced to bacteria...(protocol) At Addgene, we use the Mix & Go! E. coli Transformation Kit and Buffer Set  from Zymo Research...available via Zymo Research. In brief, we grow our E. coli in LB to log phase, then wash and resuspend the...This protocol has worked well for us for multiple E. coli strains, including DH5alpha, XL1 Blue, TOP10, ccdB... library or an especially low concentration of plasmid), you will benefit from purchasing competent cells...survivalTM, and Stbl3TM. How to transform your plasmid DNA If you decided to use commercial competent ...negative control (plate your cells without adding plasmid) to make sure you don’t have contamination.  ...

...toolbox. Natural plasmids, also called environmental plasmids, are all around us! A plasmid is a small piece...Old Solution but a New Problem Plasmids 101: What is a Plasmid? Plasmids 101: Transformation, Transduction...Gateway cloning relies on genes found on an E. coli fertility plasmid that also encodes a toxin, CcdB, and the...toxin, CcdA. Knowing which strains of E. coli contain this F plasmid is critical for successful Gateway ...tip! Historically, colicin-producing plasmids have been referred to as “Col plasmids” or “Col factors.”...F. (2023). Plasmid classifications. Plasmid, 126, 102684. https://doi.org/10.1016/j.plasmid.2023.102684...., 2001) Beneficial traits of natural plasmids Natural plasmids often convey a variety of functions that...

...on a separate plasmid. Multiplexing in E. coli Koffas Lab CRISPathBrick Multiplex Plasmid: This system ...please scroll down for plasmids suitable for other model systems, including E. coli, plants, Drosophila,...or SLIC cloning methods. A BsaI-based E. coli multiplexing plasmid is available from the Koffas lab. Scaling..., C, AB, AC, BC, and ABC). Nielsen Lab E. coli CRMAGE Plasmids: The CRMAGE system is a fast, multiplexable...specific point mutations in E. coli. gRNAs are delivered on a different plasmid and allows simultaneous multiplex...Repression in E. coli.” ACS Synthetic Biology 4(9) (2015):987-1000. PubMed PMID: 25822415. Find plasmids from ...multiple gRNAs on the same plasmid, you’ll make sure that each cell that gets the plasmid contains all of the...

...About Other Hot Plasmid Technologies Read Other Synthetic Biology Posts  Suggest Plasmid Tools for the ...Resources on Addgene.org Read our Hot Plasmids Articles Find Plasmids for Bacterial Expression ... response to changing pH, and they can rupture E. coli spheroplasts to release encapsulated proteins. ...in a single operon and were first expressed in E. coli in the 1980s  (Kanabrocki et al 1986). Much later...showed that these fluorescent R bodies can rupture E. coli spheroplasts (cells which have had their cell walls...Absence of toxicity of R-body producing recombinant E. coli on paramecia. European Journal of Protistology ...produced R-bodies that could extend at different pHs. Plasmids expressing expressing both wild type and mutant...

...resources on addgene.org Find Optogenetics Plasmids Find Biosensor Plasmids Visit the Fluorescent Protein Guide...delight,” the very first protein they tested — E. coli expressing good ol’ EGFP — showed a slight but ...1: Fluorescence intensity of EGFP-FlavinTag in E. coli changes in response to a ~25 mT magnet. Gray regions...magnetoresponse was clearly visible whether expressed in E. coli, in mammalian cells, or in purified form, and didn...license.   Video 1: Fluorescence movie of E. coli expressing MagLOV while a magnet is waved under...MagLOV plasmid here!   References and resources References Duret, G., Polali, S., Anderson, E. D., Bell...Frank Hayward et al., 2024). They deposited their plasmid with Addgene, hoping to inspire others to explore...

... knockout E. coli strain to assay for functional decaffeination in cells. The ΔguaB E. coli are missing...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...write about a recent plasmid deposit please sign up here.     Listen to the Hot Plasmids episode! Expanding.... Various soCoChR plasmids described in this work including the soCoChR-GFP plasmid as well as several...putida CBB5 and has been tailored for expression in E. coli. This operon is under the transcriptional control...Janthinobacterium sp. Moreover, a caffeine “addicted” E. coli strain expressing this operon can serve as a living...

...to find Gateway plasmids for your organisms of interest: Find Gateway cloning plasmids here! References...performed, the next step is to transform competent E. coli cells and select the positive clones. The entry... expression clone. You will also need to use a E. coli strain sensitive to CcdB (e.g. DH5α, TOP10, Mach1... protein inhibits the growth of CcdB sensitive E. coli strains, most colonies should contain the desired...available for all kind of organisms from mammals to E. coli. Glossary of Gateway cloning vectors Vector ...antibiotic resistance markers (indicated here by plasmid color), allowing you to easily select for the expression...desired, recombined construct. Read our recent Plasmids 101 Post on CcdB for more information. How to clone...

...into E. coli and plasmid assembly (re-circularization) occurs in vivo. We use XL-10 Gold E. coli, but ... assembly of a circular plasmid in vitro, before transforming it into E. coli for propagation. However...blog posts on plasmid cloning Resources on Addgene.org Read our protocols for plasmid cloning Find our...successful recombination reported in all laboratory E. coli strains tested so far. The use of this pathway ... from the MRC Laboratory of Molecular Biology. Plasmid cloning is an essential part of any molecular biology...However, while not widely known, plasmid assembly can be achieved in vivo using a bacterial recombination pathway...can be encoded in the primer sequences. Multiple plasmids can be included in a single PCR, for example for...

...tetracycline resistant plasmid, pSC101, and a newly developed kanamycin resistant plasmid, pSC102, with EcoRI...engineering, cloning, and plasmid tags, addtional content added to the "Plasmids that Glow" and "Viral Expression...Learn more about the history of plasmids and cloning: CSHL Meeting: Plasmids: History & Biology The Joshua...Lederberg in 1952 was noted to integrate into the E. coli chromosome in some cases. This terminology held...EcoRI and selected for E. coli transformants that were resistant to both. When this proved successful, ...factors played within an organism.  So how did plasmids get their name? In 1952, Joshua Lederberg set...inheritance factors. He proposed the catch-all term “plasmid” derived as a hybrid of "cytoplasm" and "id" (Latin...

...glycosylated proteins that cannot be produced in  E. coli or yeast cells. The only problem with baculovirus...bacterial host cell is almost always the answer. E. coli is definitely one of the most popular hosts for...expression may be required if your protein is toxic to E. coli. Of all the vectors, pET, under the control of ...modifications Learn more molecular biology in our Plasmids 101 eBook! Researchers use various techniques ...for your protein of interest is inserted into a plasmid expression vector that is then transformed into...being able to help the scientific community share plasmids.   Additional Resources Thermofisher Protein ...Systems Recombinant protein expression in Escherichia coli: advances and challenges Production of recombinant...

...Common Lab E. coli Strains Learn to Use Yeast Two Hybrid Systems Browse All of Our Plasmids 101 Posts ...This post is part of our ongoing Plasmids 101 series. Plasmids 101 will provide you with an overview ...of the Plasmids 101 posts here. Now that we have covered antibiotic selection here at Plasmids 101, we...α-complementation plasmid. This way you can quickly and easily identify colonies that not only have your plasmid (antibiotic...rescued by genes supplied on the transformed plasmid. Such plasmids may contain genes that allow for the use...remain the lovely whitish-cream color of standard E. coli. On the other hand, intact β-galactosidase produces...restriction digest. Make sure you use a proper E. coli strain (i.e. contains the lacZΔM15 mutation): XL1...